TM2 | Tobacco mosaic virus resistance-2
AS15 3062 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Solanum lycopersicum
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98 | 99 kDa
Brassica napus, Micromonas sp., Physcomitrella patens, Pinus sitchensis, Solanum tuberosum, Volvox carteri
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~0.5 g of Solanum lycopersicum cv M82 leaf tissue was ground with 20% TCA in acetone and finally dissolve in 200 μl UTN (urea, Tris-HCl and NaCl). The 10 μl samples were denatured with laemmli buffer at 90°C for 5 mins followed by standing on ice 2 mins. Samples were run on 10% SDS-PAGE gel and transferred to PVDF membrane by semi-dry transfer at 10V for 1 hour. Blots were then blocked with 5% non-fat milk in TBST for 2 hours at RT with agitation. Primary anti-body was diluted in 1:5000 and incubated with the blots for 2 hours at RT with agitation. Blots were rinsed twice by TBST for 15 mins each. Finally, fluorescent anti-rabbit secondary antibody was diluted in 1:50 000 and incubated with the blots for 2 hours at RT with agitation. The blots were imaged by LI-COR odyssey image system.
Courtesy Dr. Zhengming Wang, University of Cambridge, UK